Andy Powell

Two-Year Systematic Study To Assess Norovirus Contamination in Oysters from Commercial Harvesting Areas in the United Kingdom

ABSTRACT The contamination of bivalve shellfish with norovirus from human fecal sources is recognized as an important human health risk. Standardized quantitative methods for the detection of norovirus in molluscan shellfish are now available, and viral standards are being considered in the European Union and internationally. This 2-year systematic study aimed to investigate the impact of the application of these methods to the monitoring of norovirus contamination in oyster production areas in the United Kingdom. Twenty-four monthly samples of oysters from 39 United Kingdom production areas, chosen to represent a range of potential contamination risk, were tested for norovirus genogroups I and II by using a quantitative real-time reverse transcription (RT)-PCR method. Norovirus was detected in 76.2% (643/844) of samples, with all sites returning at least one positive result. Both prevalences (presence or absence) and norovirus levels varied markedly between sites. However, overall, a marked winter seasonality of contamination by both prevalence and quantity was observed. Correlations were found between norovirus contamination and potential risk indicators, including harvesting area classifications, Escherichia coli scores, and environmental temperatures. A predictive risk score for norovirus contamination was developed by using a combination of these factors. In summary, this study, the largest of its type undertaken to date, provides a systematic analysis of norovirus contamination in commercial oyster production areas in the United Kingdom. The data should assist risk managers to develop control strategies to reduce the risk of human illness resulting from norovirus contamination of bivalve molluscs.

Detection of the pufferfish toxin tetrodotoxin in European bivalves, England, 2013 to 2014.

We report the first detection of tetrodotoxins (TTX) in European bivalve shellfish. We demonstrate that TTX is present within the temperate waters of the United Kingdom, along the English Channel, and can accumulate in filter-feeding molluscs. The toxin is heat-stable and thus it cannot be eliminated during cooking. While quantified concentrations were low in comparison to published minimum lethal doses for humans, the results demonstrate that the risk to shellfish consumers should not be discarded.

Isolation of Pandemic Vibrio parahaemolyticus from UK Water and Shellfish Produce

Vibrio parahaemolyticus is a Gram-negative, halophilic bacterium found commonly in temperate and warm estuarine waters worldwide. V. parahaemolyticus is considered an emerging bacterial pathogen in Europe and has been responsible for several recent seafood-associated outbreaks. During ad hoc testing of raw shellfish produce in May 2012, pandemic group (O3:K6) V. parahaemolyticus was isolated from Pacific oysters (Crassostrea gigas), harvested in Southern England. Follow-on testing of water and shellfish, encompassing a small number geographically diverse sites, also retrieved pandemic group isolates. These strains are amongst the most northerly pandemic strains described to date and represent the first instance of pandemic V. parahaemolyticus isolated in the UK, highlighting the expanding geographical distribution of these foodborne pathogens in the environment.

Epidemiological investigation of a foodborne outbreak in Spain associated with U.S. West Coast genotypes of Vibrio parahaemolyticus

We describe an outbreak of seafood-associated Vibrio parahaemolyticus in Galicia, Spain in on 18th of August 2012 affecting 100 of the 114 passengers travelling on a food banquet cruise boat. Epidemiological information from 65 people was available from follow-on interviews, of which 51 cases showed symptoms of illness. The food items identified through the questionnaires as the most probable source of the infections was shrimp. This product was unique in showing a statistically significant and the highest OR with a value of 7.59 (1.52–37.71). All the nine strains isolated from stool samples were identified as V. parahaemolyticus, seven were positive for both virulence markers tdh and trh, a single strain was positive for trh only and the remaining strain tested negative for both trh and tdh. This is the largest foodborne Vibrio outbreak reported in Europe linked to domestically processed seafood. Moreover, this is the first instance of strains possessing both tdh+ and trh+ being implicated in an outbreak in Europe and that a combination of strains represent several pathogenicity groups and belonging to different genetic variants were isolated from a single outbreak. Clinical isolates were associated with a novel genetic variant of V. parahaemolyticus never detected before in Europe. Further analyses demonstrated that the outbreak isolates showed indistinguishable genetic profiles with hyper-virulent strains from the Pacific Northwest, USA, suggesting a recent transcontinental spread of these strains.

Fate of Human Noroviruses in Shellfish and Water Impacted by Frequent Sewage Pollution Events

Knowledge of the fate of human noroviruses (NoV) in the marine environment is key to better controlling shellfish-related NoV gastroenteritis. We quantified NoV and Escherichia coli in sewage from storm tank discharges and treated effluent processed by a UV-disinfection plant following activated sludge treatment and studied the fate of these microorganisms in an oyster harvesting area impacted by frequent stormwater discharges and infrequent freshwater discharges. Oyster monitoring sites were positioned at intervals downstream from the wastewater treatment works (WwTW) outfall impacting the harvesting area. The decay rates of NoV in oysters as a function of the distance from the outfall were less rapid than those for E. coli that had concentrations of NoV of the same order of magnitude and were over 7 km away from the outfall. Levels of E. coli in oysters from more tidally influenced areas of the estuary were higher around high water than around low water, whereas tidal flows had no influence on NoV contamination in the oysters. The study provides comparative data on the contamination profiles and loadings of NoV and E. coli in a commercial oyster fishery impacted by a WwTW.

Novel application of high pressure processing for the production of shellfish toxin matrix reference materials.

The production of homogeneous and stable matrix reference materials for marine biotoxins is important for the validation and implementation of instrumental methods of analysis. High pressure processing was investigated to ascertain potential advantages this technique may have in stabilising paralytic shellfish poisoning toxins in shellfish tissues compared to untreated materials. Oyster tissues were subjected to a range of different temperatures and pressures, with results showing a significant reduction in biological activity in comparison to control samples, without significantly altering toxin profiles. Tissue subjected to pressures >600 MPa at 50 °C was assessed for homogeneity and stability. The sample homogeneity was determined using a pre-column oxidation LC-FLD method and shown to be within accepted levels of within batch repeatability. Short and long-term stability studies were conducted over a range of temperatures, with analysis by pre and post column oxidation LC-FLD demonstrating improved stability of toxins compared to the untreated materials and with epimerisation of toxins also notably reduced in treated materials. This study confirmed the technique of high pressure processing to improve the stability of PSP toxins compared to untreated wet tissues and highlighted its applicability in reference material preparation where removal of biological activity is of importance.

Epidemic Dynamics of Vibrio parahaemolyticus Illness in a Hotspot of Disease Emergence, Galicia, Spain

Galicia in northwestern Spain has been considered a hotspot for Vibrio parahaemolyticus infections. Infections abruptly emerged in 1998 and, over the next 15 years, were associated with large outbreaks caused by strains belonging to a single clone. We report a recent transition in the epidemiologic pattern in which cases throughout the region have been linked to different and unrelated strains. Global genome-wide phylogenetic analysis revealed that most of the pathogenic strains isolated from infections were associated with globally diverse isolates, indicating frequent episodic introductions from disparate and remote sources. Moreover, we identified that the 2 major switches in the epidemic dynamics of V. parahaemolyticus in the regions, the emergence of cases and an epidemiologic shift in 2015–2016, were associated with the rise of sea surface temperature in coastal areas of Galicia. This association may represent a fundamental contributing factor in the emergence of illness linked to these introduced pathogenic strains.

Effects of denosumab on pain reduction in giant cell tumor of bone (GCTB): Interim phase II study results.

10037 Background: GCTB is a rare disease in which osteoclast-like giant cells and mononuclear cells express RANKL, a mediator of osteoclast activation. Symptoms include localized tenderness, swelling, and often severe, intractable pain. Denosumab is a fully human monoclonal antibody that binds RANKL to inhibit osteoclast-mediated bone destruction. In a previous phase 2 study, 86% of patients with GCTB responded to denosumab. This prespecified analysis of interim 12-month data from a second phase 2 study describes the effects of denosumab on pain in patients with GCTB. METHODS Adult and skeletally mature adolescent patients with GCTB (N=158) received subcutaneous denosumab 120 mg every 4 weeks with a loading dose of 120 mg SC on study days 8 and 15. The Brief Pain Inventory (BPI) Short Form (0-10) was administered before each dose. This analysis includes the 99 patients who received ≥1 dose of denosumab, were on study for ≥6 months, and completed ≥1 BPI assessment. Analyses included the proportion of patients with clinically meaningful reduction in worst pain (≥2-point decrease in patients with baseline BPI score ≥2 points), proportion of patients with no/mild worst pain at baseline who reported moderate/severe worst pain (>4 points), and change from baseline in analgesic score (0 [no analgesics]-7 [strong opioids, >600mg oral morphine equivalent/day]). RESULTS Clinically meaningful reduction (≥2-point decrease) in worst pain was reported within 1 week of their first treatment for 32% (19/60) of patients. In addition, 66% (36/55) patients had clinically meaningful reduction in worst pain at week 9. Through week 61, 22% or fewer of patients who had no/mild pain at baseline reported moderate/severe pain (>4 points). Mean analgesic score change from baseline by visit remained constant over time and ranged from -0.2 to 0.1 (median 0.0, Q1 0.0, Q3 0.0). CONCLUSIONS In this interim analysis, denosumab use in GCTB was associated with clinically meaningful reduction in worst pain in the majority of patients and the prevention of worsening pain. The reduction in worst pain and prevention of pain worsening were not accompanied by increased analgesic use.